Molecular Characterization of IL-1RAcP in Snakehead (Channa argus) and Its Immune Response to the Infection with Two Pathogenic Bacteria
LI Dong-Qi1,2, CUI Zheng-Wei3, ZHAO Fei2,*, DENG Yu-Ting2, TAN Ai-Ping2, HUANG Zhi-Bin2, JIANG Lan2
1 National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China; 2 Pearl River Fisheries Research Institute/Key Laboratory of Fishery Drug Development of Ministry of Agriculture and Rural Affairs/Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Chinese Academy of Fishery Sciences, Guangzhou 510380, China; 3 Key Laboratory of Marine Biotechnology of Fujian Province/Institute of Oceanology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Abstract Interleukin-1 receptor accessory protein (IL-1RAcP) is an important molecule involved in the pro-inflammatory cytokine interleukin-1 (IL-1) signal transduction, which plays critical roles in the inflammation, immune response and antimicrobial defense. To explore the mechanism of IL-1RAcP in the fish immune defense response to pathogen infection, the IL-1RAcP gene named shIL-1RAcP of snakehead (Channa argus) was cloned, and its immune response to the infection with 2 pathogenic bacteria and the stimulation with 3 pathogen analogues or recombinant IL-1β protein was preliminarily analyzed. The results showed that the full-length ORF of shIL-1RAcP was 1 848 bp (GenBank No. MW928496), which encoded a putative protein of 615 amino acids. Sequences analysis showed that shIL-1RAcP possessed a typical structural characterization composed of three extracellular Ig-like domains, one transmembrane domain, and one intracellular Toll/IL-1 receptor (TIR) domain. The phylogenetic tree analysis revealed that shIL-1RAcP clustered within a subgroup of other teleost species and shared the closest evolutionary relationship with the IL-1RAcP proteins of Micropterus salmoides and Seriola lalandi. Moreover, qPCR analysis showed that shIL-1RAcP was widely expressed with different expression levels in all tested tissues of healthy snakehead. The highest expression of shIL-1RAcP was in the liver, followed by several immune tissues, including the head kidney, spleen, and truck kidney, whereas the lowest expression was detected in the heart. After infection with Nocardia seriolae or Aeromonas schubertii, the relative expressions of shIL-1RAcP were mainly up-regulated in the head kidney and spleen in vivo at different time points, respectively. Similarly, after stimulation with three pathogen analogues and recombinant shIL-1β protein, the expressions of shIL-1RAcP in head kidney leukocytes in vitro could be induced by lipoteichoic acid (LTA), lipopolysaccharides (LPS), polyinosinic-polycytidylic acid [Poly (I:C)], and recombinant shIL-1β protein. In conclusion, these results illustrated that shIL-1RAcP was involved in the immune defense against the two pathogenic bacteria. Furthermore, this research provides several basic data for further revealing the mechanism of IL-1RAcP in fish species against bacterial infection.
LI Dong-Qi,CUI Zheng-Wei,ZHAO Fei, et al. Molecular Characterization of IL-1RAcP in Snakehead (Channa argus) and Its Immune Response to the Infection with Two Pathogenic Bacteria[J]. 农业生物技术学报, 2022, 30(1): 96-105.
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